Sex Pheromones of Two Leafminer Species, Antispila oinophylla and Holocacista rivillei (Lepidoptera: Heliozelidae) Infesting Grapevine in Italy.

Two heliozelid species, Antispila oinophylla van Nieukerken & Wagner and Holocacista rivillei (Stainton) severely infest Italian grapevines. The volatile pheromones from calling females were collected by solid phase micro extraction (SPME) and analyzed by gas chromatography with electroantennographic detection (GC-EAD). Two compounds from A. oinophylla females eliciting electrophysiological activity from the conspecific male antenna were identified as (Z)-5-tetradecenal and (Z)-7-tetradecenal by coupled gas chromatography/mass spectrometry (GC/MS) analysis. SPME collections from H. rivillei produced no GC-EAD active compounds but analysis of fatty acyl moieties in the pheromone gland, demonstrated the presence of the putative pheromone biosynthetic precursors (Z)-5-dodecenoic acid and (Z)-7-tetradecenoic acid. Field trapping experiments in Italy confirmed that (Z)-5-tetradecenal and (Z)-7-tetradecenal are essential for the attraction of male A. oinophylla in a blend ratio of 15:100 respectively, whereas (Z)-5-dodecenal and (Z)-7-tetradecenal attract male H. rivillei in a blend ratio of 100:6.

The impact of insecticides applied in apple orchards on the predatory mite Kampimodromus aberrans (Acari: Phytoseiidae).

Kampimodromus aberrans is an effective predatory mite in fruit orchards. The side-effects of insecticides on this species have been little studied. Field and laboratory experiments were conducted to evaluate the effects of insecticides on K. aberrans. Field experiments showed the detrimental effects of etofenprox, tau-fluvalinate and spinosad on predatory mites. Spider mite (Panonychus ulmi) populations reached higher densities on plots treated with etofenprox and tau-fluvalinate than in the other treatments. Single or multiple applications of neonicotinoids caused no detrimental effects on predatory mites. In the laboratory, spinosad and tau-fluvalinate caused 100 % mortality. Etofenprox caused a significant mortality and reduced fecundity. The remaining insecticides did not affect female survival except for imidacloprid. Thiamethoxam, clothianidin, thiacloprid, chlorpyrifos, lufenuron and methoxyfenozide were associated with a significant reduction in fecundity. No effect on fecundity was found for indoxacarb or acetamiprid. Escape rate of K. aberrans in laboratory was relatively high for etofenprox and spinosad, and to a lesser extent thiacloprid. The use of etofenprox, tau-fluvalinate and spinosad was detrimental for K. aberrans and the first two insecticides induced spider mite population increases. The remaining insecticides caused no negative effects on predatory mites in field trials. Some of them (reduced fecundity and repellence) should be considered with caution in integrated pest management programs.

Orientus ishidae (Hemiptera: Cicadellidae): Biology, Direct Damage and Preliminary Studies on Apple Proliferation Infection in Apple Orchard.

The mosaic leafhopper, Orientus ishidae (Matsumura), is an Asian species widespread in Europe that can cause leaf damage in wild trees and transmit disease phytoplasmas to grapevines. Following an O. ishidae outbreak reported in 2019 in an apple orchard in northern Italy, the biology and damage caused by this species to apples were investigated during 2020 and 2021. Our studies included observations on the O. ishidae life cycle, leaf symptoms associated to its trophic activity, and its capability to acquire "Candidatus Phytoplasma mali," a causal agent of Apple Proliferation (AP). The results indicate that O. ishidae can complete the life cycle on apple trees. Nymphs emerged between May and June, and adults were present from early July to late October, with the peak of flight between July and early August. Semi-field observations allowed for an accurate description of leaf symptoms that appeared as a distinct yellowing after a one-day exposure. In field experiments, 23% of the leaves were found damaged. In addition, 16-18% of the collected leafhoppers were found carrying AP phytoplasma. We conclude that O. ishidae has the potential to be a new apple tree pest. However, further studies are required to better understand the economic impact of the infestations.

Population dynamics of Cacopsylla melanoneura (Hemiptera: Psyllidae) in northeast Italy and its role in the apple proliferation epidemiology in apple orchards.

In the current study, incidence of 'Candidatus Phytoplasma mali' in an experimental apple orchard in northeast Italy, in addition to abundance and phytoplasma infectivity of Cacopsylla melanoneura (Förster) (Hemiptera: Psyllidae) was determined and the role of this psyllid as a vector of 'Ca. P. mali' in this region was reviewed. Insect samples collected in the orchard by the beating method indicated high abundance of C. melanoneura (up to 7.92 specimens/branch); however, the psyllid C. picta was not observed. Molecular analyses revealed presence of 'Ca. P. mali' in 6.25% of overwintered psyllids. This infection rate is quite high in comparison to other localities where C. melanoneura is known as the main vector of the phytoplasma. This finding supports the assumption that C. melanoneura also is paramount in the epidemiology of the apple proliferation disease also in northeast Italy. Moreover, we correlated immigration dynamics to the temperatures registered in the apple orchard, and defined an immigration index to predict the progressive arrival of the overwintered adults from winter sites. Psyllids start to reach the apple orchards when either the average of the maximum temperature of the 7 d is above 9.5 degrees C or the immigration index has a positive value. This index will be a useful tool for the growers to prevent apple proliferation phytoplasma spread with well-timed insecticide treatments targeted against C. melanoneura. However, further research is needed to validate or adjust the index to other apple growing regions, which may affect more efficacious management of this disease and psyllid vector.

Intraspecific Variation in Female Sex Pheromone of the Codling Moth Cydia pomonella.

The codling moth, Cydia pomonella L. (Lepidoptera, Tortricidae), is a major pest of apple, pear and walnut orchards worldwide. This pest is often controlled using the biologically friendly control method known as pheromone-based mating disruption. Mating disruption likely exerts selection on the sexual communication system of codling moth, as male and female moths will persist in their attempt to meet and mate. Surprisingly little is known on the intraspecific variation of sexual communication in this species. We started an investigation to determine the level of individual variation in the female sex pheromone composition of this moth and whether variation among different populations might be correlated with use of mating disruption against those populations. By extracting pheromone glands of individual females from a laboratory population in Canada and from populations from apple orchards in Spain and Italy, we found significant between- and within-population variation. Comparing females that had been exposed to mating disruption, or not, revealed a significant difference in sex pheromone composition for two of the minor components. Overall, the intraspecific variation observed shows the potential for a shift in female sexual signal when selection pressure is high, as is the case with continuous use of mating disruption.

Antispila oinophylla new species (Lepidoptera, Heliozelidae), a new North American grapevine leafminer invading Italian vineyards: taxonomy, DNA barcodes and life cycle.

A grapevine leafminer Antispila oinophylla van Nieukerken & Wagner, sp. n., is described both from eastern North America (type locality: Georgia) and as a new important invader in North Italian vineyards (Trentino and Veneto Region) since 2006. The species is closely related to, and previously confused with Antispila ampelopsifoliella Chambers, 1874, a species feeding on Virginia creeper Parthenocissus quinquefolia (L.) Planchon., and both are placed in an informal Antispila ampelopsifoliella group. Wing pattern, genitalia, and DNA barcode data all confirm the conspecificity of native North American populations and Italian populations. COI barcodes differ by only 0-1.23%, indicating that the Italian populations are recently established from eastern North America. The new species feeds on various wild Vitis species in North America, on cultivated Vitis vinifera L. in Italy, and also on Parthenocissus quinquefolia in Italy. North American Antispila feeding on Parthenocissus include at least two other species, one of which is Antispila ampelopsifoliella. Morphology and biology of the new species are contrasted with those of North American Antispila Hübner, 1825 species and European Holocacista rivillei (Stainton, 1855). The source population of the introduction is unknown, but cases with larvae or pupae, attached to imported plants, are a likely possibility. DNA barcodes of the three European grapevine leafminers and those of all examined Heliozelidae are highly diagnostic. North American Vitaceae-feeding Antispila form two species complexes and include several as yet unnamed taxa. The identity of three out of the four previously described North American Vitaceae-feeding species cannot be unequivocally determined without further revision, but these are held to be different from Antispila oinophylla. In Italy the biology of Antispila oinophylla was studied in a vineyard in the Trento Province (Trentino-Alto Adige Region) in 2008 and 2009. Mature larvae overwinter inside their cases, fixed to vine trunks or training stakes. The first generation flies in June. An additional generation occurs from mid-August onwards. The impact of the pest in this vineyard was significant with more than 90% of leaves infested in mid-summer. Since the initial discovery in 2006, the pest spread to several additional Italian provinces, in 2010 the incidence of infestation was locally high in commercial vineyards. Preliminary phylogenetic analyses suggest that Antispila is paraphyletic, and that the Antispila ampelopsifoliella group is related to Coptodisca Walsingham, 1895, Holocacista Walsingham & Durrant, 1909 and Antispilina Hering, 1941, all of which possess reduced wing venation. Vitaceae may be the ancestral hostplant family for modern Heliozelidae.

Effectiveness of five insecticides for the control of adults and young stages of Cacopsylla melanoneura (Förster) (Hemiptera: Psyllidae) in a semi-field trial.

BACKGROUND: Cacopsylla melanoneura (Förster) is a vector of Candidatus Phytoplasma mali, the causal agent of one of the most serious diseases in European apple orchards, apple proliferation (AP). The aim of this study was to evaluate the toxicity of five insecticides from different chemical groups (carbamates, chitin inhibitors, insect growth regulators, organophosphates and pyrethroids) towards overwintered adults and young stages of C. melanoneura under semi-field conditions. RESULTS: The organophosphate chlorpyrifos and the pyrethroid etofenprox caused higher mortality of overwintered adults than diflubenzuron, fenoxycarb and abamectin, with mortality values exceeding 90%. All the pesticides tested reduced the number of progeny per female and, except for abamectin, were highly effective against young stages, with a long-lasting effect. CONCLUSION: An integrated management of the vector is currently required to prevent the spread of AP. The reference insecticides, chlorpyrifos and etofenprox, have a strong toxicity and persistence against all stages of the vector. In apple orchard IPM programmes, the chitin inhibitors and the insect growth regulators could be part of an alternative and more sustainable strategy for the control of C. melanoneura owing to their effectiveness against immature stages.